AD

DETERMINATION OF FREE BROMINE IN WATER (U)

ANNUAL P R O G R E S S REPORT

by

T. E. Larson and F. W. S o l l o , J r .

August 1968

(For the period 1 July 1967 to 30 June 1968)

Supported by

U . S . ARMY M E D I C A L RESEARCH 6 DEVELOPMENT COMMAND

Office of The Surgeon G e n e r a l , W a s h i n g t o n , D.C. 20315
in cooperation with the Commission on Environmental Hygiene
of the Armed Forces Epidemiological Board

Contract N o . D a - 4 9 - 1 9 3 - M D - 2 9 0 9
Illinois State Water Survey
U r b a n a , Illinois 61801

DDC DISTRIBUTION STATEMENT

Each transmittal of this document outside the Department of Defense

must have prior approval of Commanding G e n e r a l , U. S. Army Medical
Research and Development Command.

The findings in this report are not to be construed as an official

Department of the Army position unless so designated by other
authorized d o c u m e n t s .
 1

I . SUMMARY

The four selected c o l o r i m e t r i c r e a g e n t s , methyl o r a n g e ,

phenol red, brom cresol p u r p l e , and p h e n o s a f r a n i n , have been
evaluated in waters containing ammonia and amino a c i d s . In
g e n e r a l , brom cresol purple and p h e n o s a f r a n i n were in
good agreement for the free b r o m i n e d e t e r m i n a t i o n s , and
methyl orange and phenol red w e r e in fairly good agreement
for the total residual bromine d e t e r m i n a t i o n s .

In a study of the disinfection of activated sludge

e f f l u e n t s , it was found that both acid o r t h o - t o l i d i n e and
methyl orange produced low results in the determination of
total available c h l o r i n e , w h e r e a s DPD gave values in better
agreement with a m p e r o m e t r i c titration v a l u e s . It appears
probable that this effect will also be found with bromine
in highly polluted w a t e r s , and for this reason DPD will be
included in future e v a l u a t i o n s .

Studies made on the stability of the phenol red, brom

cresol p u r p l e , and phenosafranin reagent s o l u t i o n s , indicate
that the three reagents are stable for four w e e k s but should
not be used t h e r e a f t e r .

Studies on the chlorination of solutions containing

ammonia and bromide ion indicate that bromamine formation
is dependent upon pH and bromide concentration and possibly
upon ammonia c o n c e n t r a t i o n . At pH 7 or a b o v e , no d e t e c t a b l e
quantity of bromamine is formed and no loss of chlorine
occurs unless the bromide ion c o n c e n t r a t i o n exceeds 5 m g / 1 .
 2

II. TABLE OF CONTENTS

Page
I SUMMARY 1
II TABLE OF CONTENTS 2
III LIST OF FIGURES 3
IV T E X T - DETERMINATION OF FREE BROMINE IN WATER
A. Introduction 4
B. Evaluation of Test M e t h o d s 5
1. Methy1 o r a n g e 5
2. Phenol red 7
3. Brom cresol purple 9
4. Phenosafranin 12
5. N , N - d i e t h y l - p - p h e n y l e n e diamine o x a l a t e 14
C. The Breakpoint Reaction for Free Chlorine and
for Free Bromine 19
D. Studies of Bromamine Solution Stability 24
E. The Reaction of C h l o r i n e with Buffered Solutions
Containing Ammonia and Bromide Ion 24
F. Bromine Demand of Ammonia and A m i n o Acids 27
G. Summary of Results 33
H. Conclusions 34
V DD FORM 1473 (DOCUMENT CONTROL DATA - R&D) 36
 3

III. L I S T OF FIGURES

FIgure Title
1 Methyl O r a n g e , Free Bromine C a l i b r a t i o n
Curve, 5 Temperatures
2 Phenol R e d , Free Bromine C a l i b r a t i o n C u r v e s ,
26°C
3 Brom Cresol P u r p l e , Free Bromine C a l i b r a t i o n
C u r v e , 26°C
4 Brom Cresol P u r p l e , Free C h l o r i n e I n t e r f e r e n c e ,
26°C
5 P h e n o s a f r a n i n , Free Bromine C a l i b r a t i o n C u r v e ,
5 Temperatures
6 DPD Oxalate R e a c t i o n Rate with C h l o r a m i n e , 26 C
7 DPD O x a l a t e , Free B r o m i n e C a l i b r a t i o n C u r v e ,
26°C
8 C h l o r a m i n e B r e a k p o i n t at pH 5.0 and pH 7 . 2 ,
26°C
9 C h l o r a m i n e B r e a k p o i n t at pH 8.1 and pH 9 . 1 ,
26°C
10 B r o m a m i n e B r e a k p o i n t at pH 5.0 and pH 7 . 3 ,
26°C
11 B r o m a m i n e B r e a k p o i n t at pH 8.4 and 9 . 2 , 26°C
12 Effect of pH on S t a b i l i t y of B r o m a m i n e S o l u t i o n ,
26°C
13 B r o m a m i n e F o r m a t i o n by C h l o r i n a t i o n of N H 3 - B r -
S o l u t i o n s , 26 C
14 B r o m i n e Demand - A m m o n i u m C h l o r i d e at pH 7.3-
8 . 0 , 26°C
15 B r o m i n e Demand - G l y c i n e at pH 6 . - 6 . 7 , 26 C
16 B r o m i n e Demand - G l y c i n e at pH 8 . - 8 . 5 , 26°C
17 B r o m i n e Demand - L-Cystine at pH 5.9-6.4, 26°C
18 B r o m i n e Demand - L-Cystine at pH 7 . 7 - 8 . 4 , 26°C
 4

IV. DETERMINATION OF FREE BROMINE IN WATER

A. Introduction

The purpose of this project is to develop a method

for the d e t e r m i n a t i o n of bromine and bromamines in w a t e r ,
with emphasis on methods that would be suitable for field
use. As previously reported, twenty-seven reagents were
evaluated for possible use as c o l o r i m e t r i c tests for free
bromine .

Four reagents were selected for detailed i n v e s t i g a t i o n ,

primarily because interferences due to c h l o r a m i n e , ferric
ion, nitrite ion, and m a n g a n e s e are low (though none is free
of all i n t e r f e r e n c e ) . The four reagents and the species
each can d e t e r m i n e a r e :

1. methyl orange - tota1 residual bromine and free

ch1orine,
2. phenol red - tota1 b r o m i n e ,
3. brom cresol purp1e - free bromine and c h l o r i n e ,
4. phenosafranin - high concentrations of free
bromine.

R e c e n t l y , the N , N - d i e t h y 1 - p - p h e n y 1 e n e diamine oxalate

colorimetric test has again been considered for the d e t e r m i n a -
tion of total residual bromine in the absence of c h l o r i n e .

The DPD o x a l a t e and the methyl orange tests have been

compared in determining the effect of pH on the stability
of bromamine solutions and show good a g r e e m e n t .

A study was made of the reaction of chlorine with

solutions containing ammonia and bromide ion. The pH
of the solutions was v a r i e d . The results indicate that a
high c o n c e n t r a t i o n of bromide ion or low pH is necessary
for the formation of b r o m a m i n e .

The four c o l o r i m e t r i c tests, brom cresol p u r p l e ,

p h e n o s a f r a n i n , phenol red, and methyl o r a n g e , h a v e been
evaluated in w a t e r containing ammonia and amino a c i d s . Free
residual bromine values determined by BCP and p h e n o s a f r a n i n
were in good a g r e e m e n t . Agreement between MO and PR tests
for total residual bromine at higher levels In the absence
of chlorine was s a t i s f a c t o r y .

In all figures the data are indicated in terms of

the halogen e m p l o y e d , except where both halogens w e r e
present In which case the results are as apparent b r o m i n e .
 5

B. EVALUATION OF TEST METHODS

All solutions were prepared using double distilled,

ammonia free w a t e r . A Sargent water bath and cooler was
used in controlled temperature s t u d i e s . Absorbance readings
were made with a Beckman Model DB spectrophotometer with
a 1.0 cm cell at the wavelength of maximum absorbance for
the reagent being tested.

1. Methyl Orange (MO)

The methyl orange test is suitable for the determina-

tion of total residual bromine in waters containing no
chlorine or total residual bromine plus free c h l o r i n e .
Addition of an excess of bromide ion to the sample
promotes a rapid quantitative reaction of chloramine with
methyl orange at p H 2 , the final pH for this method.

The calibration curves at temperatures of 2° to 40°C

are linear and cover the range of 0.0 to 4.0 mg/1 of
bromine (figure 1 ) . The MO test is slightly temperature
dependent. Ferric and nitrite ions do not interfere,
and interference due to manganic ion may be eliminated by
use of an arsenite m o d i f i c a t i o n . In the 1.5 minutes necessary
to determine bromine there is a negligible chloramine
interference. H o w e v e r , the presence of bromide ion in a
sample promotes chloramine reaction and increasing concentra-
tions of bromide ion increase the rate of reaction of c h l o r a m i n e s .

Reagents :

(1) 0.005% methyl orange is diluted from a 0.05%

stock s o l u t i o n , with 1.648 g sodium chloride
added per liter of final s o l u t i o n . This
provides a low chloride c o n c e n t r a t i o n needed
to swamp the effect of chlorides present in
the samples.

(2) 9 1 % chloroacetic acid (practical grade) -

91 g chloroacetic acid diluted to 100 ml

(3) 2.6% sodium bromide

Procedure :

To determine total residual bromine plus free c h 1 o r i n e ,

a 50 ml sample is added to 5 ml 0.005% M0 containing 1 ml
chloroacetic acid solution; this is mixed and absorbance is
determined 1.0-1.5 minutes after preparation at 505 my, pH
1.9. If the absorbance is less than 0 . 1 0 , the test is
repeated using 10 ml 0.005% methyl o r a n g e .
 Figure 1
METHYL ORANGE
FREE BROMINE CALIBRATION CURVE
1 cm CELL, X-505 my
 7

To determine tota1 residual bromine and ch1orine , 0.5

ml of 2.6% sodium bromide solution is added to the sample
after the first determination has been completed. The
solution is mixed and after 10 minutes the absorbance is
again determined. If absorbance is less than 0.10, a new
sample should be prepared using a larger quantity of methyl
orange.

2. Phenol Red (PR)

The phenol red test is suitable for the determination

of total residual bromine with no interference from free
or combined chlorine. Free bromine reacts almost
instantaneously, whereas combined bromine requires two
minutes or more for complete reaction at temperatures
in the range of 2° to 40°C.

The presence of bromamine may be detected qualitatively

in bromine solutions by adding 1 ml of 1% sodium arsenite
within 15 seconds after addition of the sample to the phenol
red in a separate test. If free bromine is the only
species present, both results will be the same. If
bromamine is present the absorbance of the second test with
arsenite will be somewhat less than that without arsenite,
due to immediate reduction of the unreacted bromamine.

The calibration curve (figure 2) for phenol red is

non-linear at low bromine concentrations. With the usual
0.01% reagent, the test is not sensitive to concentrations
of bromine less than about 0.5 mg/1. For concentrations
from 0.2 to 0.7 mg/1, a 0.001% reagent was used with some
improvement in linearity (figure 2 ) , but a cell length of
5 or 10 cm would be desirable to permit greater accuracy.

Reagents:

(1) Phenol red reagent solutions:

(a) 0.01% solution - 10 mg phenol red is
dissolved in 1 ml 0.1 N sodium hydroxide
and diluted to 100 ml. The phenol red
reagent solution (0.01%) is stable for
one month only.

(b) 0.001% solution - to 50 ml of solution

(a) is added 7 drops 0.1 N sodium hydroxide
and diluted to 500 ml.

(2) Buffer pH 5.1 - 200 ml 0.5 M sodium acetate

and 125 ml 0.4 M acetic acid.

(3) 1.0% sodium arsenite

8

Figure 2
 9

Procedure:

To determine total b r o m i n e , a 50 ml sample is added

to 2 ml phenol red reagent solution (a) or solution (b)
and 5 ml acetate buffer with m i x i n g . After 5 minutes the
absorbance is determined at 588 my, pH 5.1.

To detect b r o m a m i n e , the procedure above is repeated,

but within 15 seconds after the sample is added to the
reagent, 1 ml of 1% sodium arsenite is added. A lower
absorbance reading in this second test indicates the
presence of b r o m a m i n e .

3. Brom Cresol Purple (BCP)

The brom cresol purple calibration curve (figure 3)

is n o n - l i n e a r , particularly at concentrations below 0.50
mg/1 Br2. The curve covers the range 0.0 to 4.5 mg/1 Br2.
A new buffer having a greater buffering capacity is being
used for the brom cresol purple test.

Brom cresol purple is nearly specific for free bromine

with interference only from free c h l o r i n e . Neither
m o n o b r o m a m i n e or m o n o c h l o r a m i n e i n t e r f e r e .

At room temperature (26 ) approximately 40% of the

free chlorine present reacts with brom cresol purple within
10 m i n u t e s as shown in figure 4. The addition of sodium
arsenite within 15 seconds after the sample is added to the
reagent-buffer mixture will reduce free chlorine inter-
ference to negligible quantities if no free bromine is
present. H o w e v e r , in solutions containing free bromine
plus free chlorine the chlorine reacts with the BCP reagent
at the rate of approximately 4% for every 15 seconds
of delay before the addition of sodium a r s e n i t e . With
bromine p r e s e n t , at least 30 seconds must be allowed
before adding the arsenite to obtain full response with
the b r o m i n e . The following is an example of this:

mg/lBr2 mg/lCl2 apparent mg/1 recovered

added added as free bromine
15 sec.* 30 sec.* 60 sec.*

2.92 0.0 2.78 2.98 2.91

0.0 3.56 0.20 0.23 0.35
2.92 3.56 3.37 3.92 4.49

(3.56 mg/1 C l 2 = 8.01 mg/1 as Br2)

* Time interval before addition of arsenite

10

Figure 3
Figure 4
 12

This effect is probably due to the bromide ion in the

bromine solution which is slowly oxidized by the c h l o r i n e .

Free bromine and free c h l o r i n e together can be

determined by adding 200 mg/1 bromide ion to the sample
before testing.

Reagents;

(1) 0.0125% brom cresol p u r p l e , sodium salt-

(Dried at 103°C for 1 h r . ) . The BCP reagent
solution is stable for 1 month o n l y .

(2) Buffer solution (pH 94.) - 0.042 M borax with

6.0 ml 5N sodium h y d r o x i d e added per liter.

(3) 2.6% sodium bromide

(4 1.0% sodium arsenite

Procedure:

To d e t e r m i n e free b r o m i n e , a 50 ml sample is added

with m i x i n g , to 3 ml of 0.0125% BCP and 10 ml of buffer
solution. After 1 minute 1 ml sodium a r s e n i t e is added
and the solution is mixed w e l l . The absorbance of the
resulting solution is determined at 587 my, pH 9.3.

To d e t e r m i n e the free bromine and free chlorine

t o g e t h e r , 0.5 ml 2.6% sodium b r o m i d e solution is added
to a 50 ml sample and thoroughly m i x e d . The sample is
then added, with m i x i n g , to 3 ml 0 . 0 1 2 5 % BCP and 10 ml
b u f f e r , and after 1 minute the absorbance is determined
at 587 mu, as b e f o r e .

4. Phenosafranin

The phenosafranin test may be used for the d e t e r m i n a -

tion of free b r o m i n e . The reaction between free b r o m i n e
and p h e n o s a f r a n i n does not appear to be temperature
dependent in the range 2° to 40°C (figure 5 ) . The curve
covers the range of 0.0 to 10 mg/1 B r 2 . Because of its
rather low sensitivity with a 1 cm c e l l , this reagent
has 1imited v a l u e .

B r o m a m i n e and free chlorine react slowly with

p h e n o s a f r a n i n but the addition of 1 ml of 1% sodium
a r s e n i t e within 15 seconds after the sample is added to
the reagent will eliminate both of these i n t e r f e r e n c e s .
13

Figure 5
 14

Reagents ;

(1) 0.01% phenosafranin

The phenosafranin reagent solution is
stable only for 1 month.

(2) Buffer, pH 9.2 - 0.042 M borax

(3) 1.0% sodium arsenite

Procedure ;

To determine free bromine, a 50 ml sample is added

with mixing to 3 ml of 0.01% phenosafranin and 5 ml
buffer. After 15 seconds, 1 ml 1% sodium arsenite is
added. Absorbance is determined within 1 minute at
520 my, pH 9.2.

5. N,N-diethy1-p-pheny1ene diamine oxalate (DPP oxalate)

Two procedures were evaluated: a colorimetric

procedure as used in this laboratory for chlorine determina-
tion 1 and Palin's ferrous ammonium sulfate (FAS) titration
procedure 2 . The colorimetric test was judged to be the
more accurate procedure. With either test, chloramine
interferes to some extent (figure 6 ) . Chloramine inter-
ference was greater with the titration procedure; during
the one minute required to complete the titration, about
10% of the chloramine present reacted with the DPD, and
the continuing chloramine-DPD reaction masked an already
indistinct endpoint.

Both free bromine and bromamine react rapidly with

DPD. Free bromine reacts within 1 minute, while about
1.5 minutes should be allowed to insure complete bromamine
reaction. Chlorine and bromine concentration may be
evaluated separately by the addition of ammonium sulfate
to the sample to form halogen amines 1 minute before
addition of the samples to the reagent. Deleting the
ammonium sulfate step allows free chlorine, free bromine,
and bromamine to be evaluated free of chloramine; and
addition of potassium iodide promotes the reaction of
chloramine to give total halogen.

1. A. T. Palin, "The Determination of Free and Combined

Chlorine In Water by the Use of Diethyl-p-phenylene
Diamine," J.AWWA, Vol. 49. pp.873-880 (July 1957).
2. A. T. Palin, "The Determination of Free Residual Bromine
in Water," Water and Sewage Works, Vol. 108, p.461 (1961).
 15

In previous work on chlorine it was established that

+3
Mn interferes with the test, but that F e + 3 and NO do n o t 3 .
An arsenite modification should be used to eliminate M n + 3
interference (0.25 ml 1% sodium arsenite per 100 ml s a m p l e ) .
The reagent as used for chlorine is not temperature
dependent, and it is assumed that the same would be true
for the bromine test. The free bromine calibration curve
(figure 7) is linear from 0.0 to 2.5 mg/1 B r 2 but with
an appropriate calibration the range may be extended to
8 mg/1 .

Reagents I

Colorimetric test -
(1) OPD oxalate indicator solution: 1 g DPD
oxalate is dissolved in double distilled
water containing 8 ml 1+3 sulfuric acid
and 0.2 m disodium ethy1enediamine tetra-
acetate dihydrate ( E D T A ) . This is diluted
to 1 liter, stored in an amber glass-stoppered
b o t t l e . The reagent, if stored in a refrigerator,
can be used for one m o n t h .

(2) Buffer (pH 6.5) - 24 g dibasic sodium phosphate

and 46 g monobasic potassium phosphate are
dissolved in double distilled water. This
solution is combined with 100 ml d.d. water
in which 0.8 g EDTA has been dissolved, and
then diluted to 1 liter and 20 mg mercuric
chloride added .

(3) 1.0% sodium arsenite

(4) Ammonium sulfate (analytical reagent)
Procedure I

Colorimetric test -
(a) To determine tota1 b r o m i n e , 50 mg ammonium sulfate
is added to 100 ml of s a m p l e , m i x e d , and allowed to stand
for 1 m i n u t e . This solution is then added to a mixture of
5 ml DPD indicator and 5 ml buffer and mixed thoroughly.
The absorbance is determined in 1.5 minutes at 552 m y , pH 6.2.

3. T. E. Larson, F. W. S o l l o , "Determination of Free

Chlorine Residuals in W a t e r , " Final Technical Report to
the Commission on Environmental Hygiene of the Armed Forces
Epidemiological Board, Contract N o . D A - 4 9 - 1 9 3 - M D - 2 3 9 9 ,
15 February 1963 to 31 August 1965, p . 8 .
 16

(b) To determine free ch1orine plus t o t a l b r o m i n e ,

100 ml of sample is added to a mixture of 5 ml DPD indicator
and 5 ml buffer and mixed thoroughly. The absorbance is
determined in 1.5 m i n u t e s at 552 mµ, pH 6.2

(c) To determine total ha1ogen (total b r o m i n e + total

c h l o r i n e ) , 100 ml of sample is added to a mixture of 5 ml
DPD indicator and 5 ml b u f f e r . Then 1 gram potassium
iodide is added, and the absorbance is determined in 5
minutes at 552 my, pH 6.2.

Reagents II

FAS titration:

(1) DPD indicator solution as in c o l o r i m e t r i c test

(2) Buffer (pH 6.5) as in colorimetric test
(3) Standard ferrous ammonium sulfate titrant -
1.106 g ferrous ammonium sulfate (Mohr's salt)
is dissolved in double distilled water containing
1 ml 1+3 sulfuric acid and diluted to 1 liter
with freshly boiled and cooled distilled water.
The reagent is standardized against 0.0025N
standard potassium d i c h r o m a t e .
(4) Potassium iodide (analytical reagent)
(5) Ammonium sulfate (analytical reagent)
(6) Ferroin indicator - 1.485 g o-phenan throline
m o n o h y d r a t e and 0.695 g ferrous sulfate is
dissolved in double distilled w a t e r and
d i luted to 100 m l .

Procedure II

FAS titration:

(a) To determine tota1 b r o m i n e , 50 mg ammonium

sulfate is added to 100 ml of s a m p l e , m i x e d , and allowed
to stand for 1 m i n u t e . Then the sample is added to a
mixture of 5 ml DPD indicator and 5 ml buffer and titrated
with FAS to a colorless solution.

(b) To determine tota1 bromine + free c h l o r i n e ,

100 ml of sample is added to a mixture of 5 ml DPD indicator
and 5 ml b u f f e r , and then titrated with FAS to a colorless
solution.

(c) To determine tota1 ha1ogen (total bromine + total

c h l o r i n e ) , 100 ml of samp1e is added to a m i x t u r e of 5 ml
DPD indicator and 5 ml b u f f e r . Then 1 gram potassium iodide
is added and after 2 minutes the sample is titrated with FAS
to a colorless s o l u t i o n .
Figure 6
 Figure 7
DPD OXALATE
. FREE BROMINE CALIBRATION CURVE
1 cm CELL X-552 my
ROOM TEMPERATURE (26°C)
 19

C. THE BREAKPOINT REACTION FOR FREE CHLORINE

AND FOR FREE BROMINE

Curves showing chlorine and bromine breakpoint

phenomena for w a t e r with an ammonia-nitrogen concentra-
tion of 1 mg/1 and reaction time of one hour have been
prepared at pH 5.0, 7.2, 8.4, and 9.1 (figures 8 to 1 1 ) .

On a molar b a s i s , the ammonia-bromine reaction

appears to be similar to the ammonia-chlorine reaction.
The bromamine breakpoint requires 14.1 mg/1 of free
b r o m i n e , while the chloramine breakpoint requires 6.3
mg/1 of free c h l o r i n e .

As reported in our Progress Report of August 1967

and as can be observed in figures 8 to 11 of this report,
amine stability is better for chlorine than for b r o m i n e .

Methly o r a n g e , neutral o r t h o t o 1 i d i n e , and DPD were

the tests used in preparing the ch1orine breakpoint
curves as shown in figures 8 and 9. Each of these tests
was used to determine both free and total c h l o r i n e . Agree-
ment among the tests for the determination of free chlorine
was poorest at pH 9.1. Neutral o r t h o t o 1 i d i n e and DPD
values are considerably higher than methyl orange values
for free c h l o r i n e . Repeated work at this pH gave the
same r e s u l t s . It appears that free chlorine and chloramine
coexist for a longer time at pH 9.1, and the breakpoint
is not distinct under these c o n d i t i o n s . The lower pH of
the test conditions for the MO determination promotes
excessive reaction between the free and combined c h l o r i n e .

Brom cresol p u r p l e , phenosafran in, phenol red, methyl

o r a n g e , and DPD w e r e the tests used for preparing the
bromine breakpoint c u r v e s . BCP and phenosafranin were
used to d e t e r m i n e free b r o m i n e ; PR, M O , and DPD were used
for the d e t e r m i n a t i o n of total bromine (figures 10 and 1 1 ) .

Samples w e r e prepared by the addition of the desired

concentration of free B r 2 or C 1 2 to a buffered solution
containing 1 mg/1 NH3 while mixing with a magnetic s t i r r e r .

4. J. Donald J o h n s o n . "Development of Field Test for Free

Chlorine," Annual Progress Report to the Commission on
Environmental Hygiene of the Armed Forces Epidemiological
Board, Contract No. D A - 4 9 1 9 3 - M D - 2 4 4 2 , 1 March 1966 to
30 June 1 9 6 7 .
 20

Figure 8
21

Figure 9
22

Figure 10
23

Figure 11
 24

The samples were stored in glass-stoppered bottles and aliquots

extracted after one hour for testing. Tests were performed as
nearly simultaneously as p o s s i b l e , but there was a time lapse
of from 3 to 5 minutes from the addition of the sample for the
first test to the addition of the sample for the fifth test.
This may account for some variation in results with different
tests.

D. STUDIES OF BROMAMINE STABILITY

A series of stability tests was carried out at room

temperature with 1000 mg/1 NH 3 + 50 mg/1 B r 2 and at pH values
of 5.95, 7.1, 7.8, and 8.8 (figure 1 2 ) . Since the DPD oxalate
test is once again being considered for total bromine determina-
tion, the sole purpose of this series of b r o m a m i n e stability
work was to make a comparison of the methyl orange test results
with the DPD test results. The two tests were in good agree-
ment in the determination of total b r o m i n e .

The test procedure w a s as f o l l o w s : Free bromine (as NaOBr)

was added to a buffered solution containing 1000 mg/1 ammonia.
During bromine a d d i t i o n , samples w e r e mixed by means of a
magnetic stirrer to insure complete mixing and to e l i m i n a t e the
possibility of error due to local e x c e s s e s of bromine or
ammonia. At specified time i n t e r v a l s , samples were withdrawn
and analyzed for total bromine using the DPD and the methyl
orange p r o c e d u r e s .

E. THE REACTION OF CHLORINE WITH BUFFERED SOLUTIONS

CONTAINING AMMONIA AND BROMIDE ION

A study was made of the chemistry of the reactions of

chlorine when added to solutions containing ammonia and bromide
ion and at varying pH (figure 1 3 ) .

1.85 mg/1 free chlorine was added to previously buffered

samples containing 1 mg/1 ammonia (from ammonium chloride) and
bromide ion (from sodium b r o m i d e ) . The bromide ion concentra-
tions w e r e 1, 3, 5, 2 5 , 1 0 0 , and 250 m g / 1 . The range of pH
values was from 5 to 9. The total bromine c o n c e n t r a t i o n s w e r e
determined after 10 m i n u t e s contact time by the phenol red test.
The phenol red test was used because it is completely insensitive
to chlorine and c h l o r a m i n e s . Ultra violet absorption was used
to determine the species of halogen-amine that was formed.

An examination of figure 13 shows that b r o m a m i n e formation

is dependent upon pH and bromide ion c o n c e n t r a t i o n . At pH 7 or
a b o v e , no detectable quantity of bromamine is formed unless the
bromide ion concentration exceeds 5 m g / 1 .

Total available halogen was also determined with methyl

orange. With the higher bromide c o n c e n t r a t i o n s , methyl orange
values were lower than the phenol red r e s u l t s , due to decomposi-
tion of a portion of the bromamines at the low pH required in
the methyl orange test.
 25

EFFECT OF pH ON STABILITY OF BROMAMINE SOLUTION

AT ROOM TEMPERATURE (26°C)

Figure 12
 Initial conditions:

Residual Bromine was

determined by Phenol
Red test after 10 min.
contact time

Figure 13
 27

In the presence of 25 mg/1 of bromide ion or m o r e ,

the formation of some bromamine occurs over the entire
pH range tested. T h e r e f o r e , it appears that a high concentra-
tion of bromide ion or low pH is necessary for the bromide
ion to be oxidized and subsequently form b r o m a m i n e .

Ultra violet absorption showed that under the reaction

conditions stated in paragraph 2 a b o v e , tribromamine is
formed at pH values between pH5 and p H 6 , a mixture of
tribromamine and m o n o c h l o r a m i n e is formed at pH values
between 6 and 7» and m o n o c h 1 o r a m i n e alone is formed at
pH values between pH7 and p H 9 .

Work is in progress to determine the effect of varying

c o n c e n t r a t i o n s of ammonia on this reaction.

F. BROMINE DEMAND OF AMMONIA AND AMINO ACIDS

The four colorimetric procedures for the determina-

tion of free and combined bromine were evaluated for
their performance in water containing ammonia and amino
acids. The tests under study are brom cresol purple
and phenosafranin for free bromine and methyl orange
and phenol red for total available b r o m i n e .

Varying c o n c e n t r a t i o n s of bromine were added to

ammonia or amino acid samples and after the desired
contact times free and total residual bromine were
determined by the proper tests.

The water used for these tests was free of ammonia

and contained 150 mg/1 each of alkalinity and h a r d n e s s .
The pH was adjusted with carbon d i o x i d e .

Solutions of ammonium c h l o r i d e , g l y c i n e , and L-cystine

were prepared in ammonia-free water containing 150 mg per
liter each of alkalinity and hardness (prepared as described
above) and adjusted to pH 6, 7, or 8. For each sample the
nitrogen (N) c o n c e n t r a t i o n was 0.3 m g / 1 . The temperature
of the samples and the tests was 26 C (room t e m p e r a t u r e ) .
The contact times were 1 hr., 4 h r s . , and 2k h r s .

The ammonium chloride samples were prepared only at

pH 7 (figure 14). The breakpoint for these samples occurred
at a free bromine dosage of 6.2 mg/1 (as B r 2 ) . Brom cresol
purple and p h e n o s a f r a n i n were in good agreement for free
residual bromine as were the methyl orange and phenol red
values for tota1 residual b r o m i n e .
 28

BROMINE DEMAND - AMMONIUM CHLORIDE (0.3 mg/1 N ) : pH 7.3-8, 26°C

150 mg/1 each ALKALINITY AND HARDNESS

Figure 14
 29

BROMINE DEMAND - GLYCINE (0.3 mg/1 N ) : pH 6-6.7, 26°C,

150 mg/1 each ALKALINITY AND HARDNESS

Figure 15
 30

BROMINE DEMAND - GLYCINE (0.3 mg/1 N): pH 8 - 8 . 5 , 26°C

150 mg/1 each ALKALINITY AND HARDNESS

Figure 16
 31

BROMINE DEMAND - L-CYSTINE (0.3 mg/1 N ) : pH 5.9-6.4, 26°C

150 mg/1 each ALKALINITY AND HARDNESS

Figure 17
 32

BROMINE DEMAND - L-CYSTINE (0.3 mg/1 N ) ; pH 7.7-8.4, 26°C

150 mg/1 each ALKALINITY AND HARDNESS

Figure 18
 33

G 1 y c i n e - b r o m i n e samples were prepared at pH values

of 6.5 and 8, (figures 15 and 1 6 ) . The glycine concentra-
tion was equivalent to 0.3 mg/1 of n i t r o g e n . Samples at
both pH values showed a breakpoint at a dosage of about
12 mg/1 free b r o m i n e . Prior to the b r e a k p o i n t , the methyl
orange values were higher than the phenol red values for
total residual b r o m i n e .

L-cystine was treated with bromine at pH values of

6 and 8, (figures 17 and 1 8 ) . Methyl orange values were
considerably higher than phenol red values for total b r o m i n e .
Such apparent differences in results for total bromine
are probably due to the non-linearity of the phenol red
calibration curve with low B r 2 c o n c e n t r a t i o n s .

The brom cresol purple and the phenol red tests were
modified slightly in order to improve r e s u l t s . For the
brom cresol purple test, the pH was raised from 9 to 9.30
by using 10 ml pH 9.4 borate-sodiurm hydroxide buffer. In
order to determine the very low total bromine residuals
more accurately (0-0.7 mg/1 Br 2 ) a phenol red reagent
solution of 0.001% strength was used.

In general the values for free bromine determined by

BCP (pH 9.3) and phenosafranin (pH 9.1) were in good agree-
ment. The methyl orange test (pH 2) showed higher total
residual bromine values prior to the breakpoint than did
the phenol red test (pH 5 . 1 ) .

A n o t h e r project in this laboratory deals with the

disinfection of sewage e f f l u e n t s . In this w o r k , it was
found that the amperometric titration and the DPD test
gave results which were in good a g r e e m e n t , w h e r e a s the
o r t h o t o l i d i n e and methyl orange procedures gave much lower
results. P r e s u m a b l y , this is due to a rapid reaction of
the chloramines with reducing substances in the effluents
at the low pH (1 and 2) required for the t e s t s .

It is assumed that a similar effect would be observed

with b r o m i n e , and tests are planned to verify t h i s .

G. SUMMARY OF RESULTS

Since the last Annual Progress Report, the four

c o l o r i m e t r i c tests recommended for the determination of
free bromine and bromamine , brom cresol p u r p l e , pheno-
s a f r a n i n , methyl o r a n g e , and phenol red, have been further
evaluated. More recent work has included Palin's DPD
o x a l a t e test for total " r e s i d u a l " b r o m i n e . M i n o r changes
in reagent concentration and buffer strength for PR and BCP
 34

have become necessary as work has p r o g r e s s e d . The

reagents have been tested for stability which appears to
be one month for PR, BCP, phenosafranin and DPD. MO is
stable for at least three m o n t h s .

The reactions of free chlorine in buffered solutions

containing 1 mg/1 ammonia nitrogen and varying concentra-
tions of bromide ion have been studied in greater d e t a i l .
Results indicate that high c o n c e n t r a t i o n s of bromide ion
or low pH are necessary for formation of b r o m a m i n e s .

Some bromamine stability studies for one hour contact

times were repeated for the purpose of comparing MO and
DPD test results. The results are in good agreement.

An evaluation of four c o l o r i m e t r i c t e s t s , brom cresol

p u r p l e , p h e n o s a f r a n i n , methyl o r a n g e , and phenol red is
being made in water containing ammonia and amino a c i d s .
Brom cresol purple and phenosafranin show good agreement
in the d e t e r m i n a t i o n of free b r o m i n e , while methyl orange
and phenol red are in fairly good agreement in the d e t e r m i n a -
tion of total b r o m i n e . This work is a preliminary to an
evaluation of the p e r f o r m a n c e of these tests in polluted
water. To such an evaluation will be added a fifth test,
DPD, for total b r o m i n e . The DPD test will be used since
other work in this laboratory has shown that at the low
pH required by the MO test there may be a rapid reaction
of the chloramines (and possibly the b r o m a m i n e s , though
this has not yet been tested) with reducing substances in
the sample.

H. CONCLUSIONS

The brom cresol purple and p h e n o s a f r a n i n tests are

suitable for the d e t e r m i n a t i o n of free b r o m i n e . The DPD
o x a l a t e , methyl o r a n g e , and phenol red tests are suitable
for the d e t e r m i n a t i o n of tota1 b r o m i n e .

Studies of the bromine demand of solutions of ammonia

and amino acids indicate good agreement in results among
the tests for free bromine and b r o m a m i n e .

Studies to date of the reaction of chlorine with

solutions containing bromide
in order to form b r o m a m i n e s
of chlorine and bromide ion,
to a small volume at low pH
mixing of this solution with
treated.
ion and ammonia indicate that
in a w a t e r supply by addition
these would have to be added
for bromine formation with subsequent
the remainder of the water to be
 35

Future work will include a more thorough evaluation

of the DPD oxalate test for the determination of total
bromine. The effect of temperature and interfering
substances will be determined.

The DPD test as well as the brom cresol p u r p l e ,

p h e n o s a f r a n i n , methyl o r a n g e , and phenol red tests will
be evaluated for their performance in highly polluted w a t e r .

Future work will include a more detailed study of

the reaction of chlorine with solutions containing ammonia
and bromide ion, and in p a r t i c u l a r , the effect of varying
c o n c e n t r a t i o n s of ammonia in such s o l u t i o n s .
 36
Security Classification
DOCUMENT CONTROL DATA - R&D
(Security claasification of (title, body of absfract and Indenting annotation must be entered when the overall report la classified)
1 ORIGINATING ACTIVITY (Corporate author) 2a. REPORT SECURITY CLASSIFICATION

ILLINOIS STATE WATER SURVEY 2b OROUP

Urbana, Illionis
3 REPORT T I T L E

DETERMINATION OF FREE BROMINE IN WATER

4 DESCRIPTIVE NOTES (Typo of report and incluaive dates)

Annual Progress Report 1 July 1967 to 30 June 1968
5 AUTHOR(S) (Last name. first name, Initial)

T. E. Larson and F. W. Sollo, Jr.

6 REPORT DATE 7a. TOTAL NO OF PACES 7b. NO. OP REPS

August 1968 38
8a CONTRACT OR ORANT NO. 9a. ORIGINATOR'S REPORT NUMBER(S)

DA-49-193-MD-2909 2
b. PROJECT NO.

c. 9b. OTHER REPORT NO(S) (Any other numbers that may be resigned
this report)

d
1 0 A V A I L A B I L I T Y / L I M I T A T I O N NOTICES

Qualified requesters may obtain copies of this report from DDC.

11 SUPPLEMENTARY NOTES 12. SPONSORING MILITARY A C T I V I T Y

U . S . Army M e d i c a l R e s e a r c h and
D e v e l o p m e n t Command, W a s h i n g t o n , D.C.
13 ABSTRACT 20315

The f o u r s e l e c t e d c o l o r i m e t r i c r e a g e n t s , methyl o r a n g e , phenol

r e d , b r o m c r e s o l p u r p l e , and p h e n o s a f r a n i n , h a v e b e e n e v a l u a t e d i n
w a t e r s c o n t a i n i n g a m m o n i a and a m i n o a c i d s . I n g e n e r a l , brom c r e s o l
p u r p l e and p h e n o s a f r a n i n w e r e i n g o o d a g r e e m e n t f o r t h e f r e e b r o m i n e
d e t e r m i n a t i o n s , and m e t h y l o r a n g e and p h e n o l r e d w e r e i n f a i r l y
good agreement f o r t h e t o t a l r e s i d u a l b r o m i n e d e t e r m i n a t i o n s .

In a study of the d i s i n f e c t i o n of a c t i v a t e d sludge e f f l u e n t s , it

was f o u n d t h a t b o t h a c i d o r t h o - t o l i d i n e and m e t h y l o r a n g e p r o d u c e d
low r e s u l t s i n t h e d e t e r m i n a t i o n o f t o t a l a v a i l a b l e c h l o r i n e , whereas
DPD g a v e v a l u e s i n b e t t e r a g r e e m e n t w i t h a m p e r o m e t r i c t i t r a t i o n
values. It appears probable t h a t t h i s e f f e c t w i l l a l s o be found
w i t h b r o m i n e i n h i g h l y p o l l u t e d w a t e r s , and f o r t h i s r e a s o n DPD w i l l
be included in f u t u r e e v a l u a t i o n s .

S t u d i e s made o n t h e s t a b i l i t y o f t h e p h e n o l r e d , b r o m c r e s o l
p u r p l e , and p h e n o s a f r a n i n r e a g e n t s o l u t i o n s i n d i c a t e t h a t t h e t h r e e
r e a g e n t s are s t a b l e f o r f o u r weeks but s h o u l d n o t b e used t h e r e a f t e r .

DD 1473
 37

14. KEY W O R D S
Analysis
Water
Bromine
Methyl O r a n g e
Phenol Red
Brom Cresol P u r p l e
Phenosafranin
N,N-diethyl-p-pheny1ene diamine Oxalate
Abstract (cont'd) 38

Studies on the chlorination of solutions containing

ammonia and bromide ion indicate that bromamine formation
is dependent upon pH and bromide concentration and possibly
upon ammonia c o n c e n t r a t i o n . At pH 7 or a b o v e , no detectable
quantity of bromamine is formed and no loss of chlorine
occurs unless the bromide ion concentration exceeds 5 m g / 1 .